Fermentation method for improving yield of bacillus subtilis antibacterial protein

A technology of Bacillus subtilis and Bacillus subtilis is applied in the field of fermentation to improve the yield of Bacillus subtilis antibacterial protein, and can solve the problems of immature industrial application, incomplete promotion of drugs, unclear characteristics and mechanism of action, etc., and achieve stable yield , the effect of saving costs and improving production levels

Inactive Publication Date: 2019-01-08
SHAANXI UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although there are still many problems in the application of Bacillus subtilis, such as immature industrial applicat...

Method used

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  • Fermentation method for improving yield of bacillus subtilis antibacterial protein
  • Fermentation method for improving yield of bacillus subtilis antibacterial protein
  • Fermentation method for improving yield of bacillus subtilis antibacterial protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] S1. Purchase the strain of Bacillus subtilis, the number of which is BNCC 189983, which is preserved by BNCC Biotechnology Co., Ltd. (BNCC). Inoculate the Bacillus subtilis strain on the slant and cultivate it in a constant temperature incubator at 37°C, then prepare a bacterial suspension with a concentration of 5,000,000 cells / ml from the activated strain on the slant medium, and inoculate it in a fresh In the seed medium of 36°C, shake culture on a shaker at 180r / min.

[0043] S2. Inoculate the fermented strain BNCC 189983 Bacillus subtilis to nutrient components (according to mass percentage) as glucose 1.0%, peptone 1.0%, NaCl 1.0%, KH 2 PO 4 0.15%, MgSO 4 ·7H 2 O 0.15%, the rest of the water in the seed medium was cultivated for 12 hours to obtain a seed liquid, the culture temperature was 36° C., and the initial pH value of the medium was 7.0.

[0044] S3. Inoculate the seed solution of the strain producing the Bacillus subtilis BNCC 189983 antibacterial pro...

Embodiment 2

[0052] S1. Purchase the strain of Bacillus subtilis, the number of which is BNCC 189983, which is preserved by BNCC Biotechnology Co., Ltd. (BNCC). Inoculate the Bacillus subtilis strain on the slant and cultivate it in a constant temperature incubator at 37°C, then prepare a bacterial suspension with a concentration of 5,000,000 cells / ml from the activated strain on the slant medium, and inoculate it in a fresh In the seed medium of 37 ° C, 200 r / min shaking on the shaker culture.

[0053] S2. Inoculate the fermented strain BNCC 189983 Bacillus subtilis to nutrient components (according to mass percentage) as glucose 1.0%, peptone 1.0%, NaCl 1.0%, KH 2 PO 4 0.15%, MgSO 4 ·7H 2 O 0.15%, the rest of the water in the seed medium was cultivated for 14 hours to obtain a seed solution, the culture temperature was 37° C., and the initial pH value of the medium was 7.0.

[0054] S3. Inoculate the seed liquid of the strain producing the Bacillus subtilis BNCC 189983 antibacterial...

Embodiment 3

[0062] S1. Purchase the strain of Bacillus subtilis, the number of which is BNCC 189983, which is preserved by BNCC Biotechnology Co., Ltd. (BNCC). Inoculate the Bacillus subtilis strain on the slant and cultivate it in a constant temperature incubator at 37°C, then prepare a bacterial suspension with a concentration of 5,000,000 cells / ml from the activated strain on the slant medium, and inoculate it in a fresh In the seed medium of 38 ° C, 220 r / min shaking on the shaker culture.

[0063] S2. Inoculate the fermented strain BNCC 189983 Bacillus subtilis to nutrient components (according to mass percentage) as glucose 1.0%, peptone 1.0%, NaCl 1.0%, KH 2 PO 4 0.15%, MgSO 4 ·7H 2 O 0.15%, the rest of the water in the seed medium was cultivated for 16 hours to obtain a seed solution, the culture temperature was 37° C., and the initial pH value of the medium was 7.0.

[0064] S3. Inoculate the seed liquid of the bacteria producing the Bacillus subtilis BNCC 189983 antibacteri...

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Abstract

The invention discloses a fermentation method for improving the yield of bacillus subtilis antibacterial protein. The method comprises the following steps: firstly obtaining microbial strain, successively inoculating the obtained microbial strain to a seed culture medium and a fermentation culture medium to perform fermentation, so as to obtain the bacillus subtilis antibacterial protein, and performing stage adjustment according to growth conditions of thallus in a logarithmic growth phase within 6 to 16 hours after the beginning of fermentation of the thallus and in a stable phase from 16 hours after the beginning of fermentation of the thallus to the end of the fermentation and according to the fermentation time. The method has the advantages that the inoculation amount of the microbialstrain is regulated before the fermentation of the bacillus subtilis BNCC 189983 antibacterial protein, the temperature, dissolved oxygen, pH value and shaking table rotate speed of a system during fermentation are regulated, the microbial strain can rapidly and stably grow in the early stage of fermentation through the regulation of the parameters, the microbial strain can efficiently and stablyproduce antibacterial protein in the middle and later stage, and the production of other side products is reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a fermentation method for increasing the yield of antibacterial protein of Bacillus subtilis. Background technique [0002] Bacillus, as an important class of biocontrol bacteria, can produce various antibacterial and antibacterial substances to antagonize plant pathogenic bacteria, has the potential to prevent and control plant diseases, and is also the main source of biological control. [0003] Among them, Bacillus subtilis is a non-pathogenic bacillus that exists widely in nature, is non-toxic and harmless to humans and animals, does not pollute the environment, and has inhibitory effects on plant diseases. At present, many researchers have extensively studied the pathogenic antagonism of Bacillus subtilis and other bacillus and their antibacterial substances. Bacillus can produce a variety of antibacterial substances to prevent and control diseases, including bacter...

Claims

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Application Information

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IPC IPC(8): C12P21/00C12N1/20C12R1/125
CPCC12N1/20C12P21/00
Inventor 龚国利刘志强张曦文佐静
Owner SHAANXI UNIV OF SCI & TECH
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